Faculty of Biology, University of Latvia, 4 Kronvalda Boulevard, 1586 Riga, Latvia.
J Anal Methods Chem. 2012;2012:103575. doi: 10.1155/2012/103575. Epub 2012 Apr 3.
A simple, sensitive, precise, and specific reverse HPLC method was developed and validated for the determination of plant hormones in barley (Hordeum vulgare L.). The method includes extraction in aqueous organic solvent followed by solid-phase extraction, sample evaporation, and reversed-phase HPLC analysis in a general purpose UV-visible (abscisic acid (ABA)) and fluorescence detection (indole-3-acetic acid (IAA) and indole-3-pyruvic acid (IPA)), high-performance liquid chromatography system. The separation was carried out on Zorbax Eclipse XDB C8 column (150 × 4.6 mm I.D) with a mobile phase composed of methanol and 1% acetic acid (60 : 40 v/v) in isocratic mode at a flow rate of 1 ml min(-1). The detection was monitored at 270 nm (ABA) and at 282 nm (Ex) and 360 nm (Em) (IAA, IPA). The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantification, and robustness. The determined validation parameters are in the commonly acceptable ranges for that kind of analysis.
建立并验证了一种用于大麦中植物激素检测的简单、灵敏、准确、特异的反相高效液相色谱法。该方法包括在水-有机溶剂中提取，然后固相萃取、蒸发、反向高效液相色谱分析（用紫外-可见（脱落酸（ABA））和荧光检测（吲哚-3-乙酸（IAA）和吲哚-3-丙酮酸（IPA）））。在一般用途的高效液相色谱系统中进行。分离是在 Zorbax Eclipse XDB C8 柱（150 × 4.6 毫米内径）上进行的，流动相由甲醇和 1%乙酸（60：40 v/v）组成，采用等度洗脱方式，流速为 1ml/min。检测在 270nm（ABA）和 282nm（Ex）和 360nm（Em）（IAA、IPA）处进行。该方法在准确度、精密度、线性、检测限、定量限和稳健性方面进行了验证。所确定的验证参数在该类分析中通常可接受的范围内。
